Difference between revisions of "Part:BBa K3505024"
Line 12: Line 12:
 
===Design Notes===
 
===Design Notes===
 
The coding sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is presents in pUPD2 and has overhangs compatible for Golden Braid cloning.  
 
The coding sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is presents in pUPD2 and has overhangs compatible for Golden Braid cloning.  
−
The CDS has position B3-B5.
+
The CDS has position B2-B5.
 +
 
 +
<p style="text-align: center;">
 +
    [[Image: T--Thessaly--GB-CCAT-GCTT.jpeg .jpeg|900px|thumb|none|<i><b>Figure 1.</b>The overhangs of this part in the Golden Braid Grammar.</i>]]
 +
 
 +
 
 +
[[Image:T--Thessaly--b-arrestin-snap.png|900px|thumb|none|<I><b>Figure 2.</b> The level 0 module : pupd2-b-arrestin-TEV protease (illustration from SnapGene)</i>]]
 +
 
 +
===Verification of Cloning===
 +
[[File:T--Thessaly--b-arrestin-digestion.png|700px|thumb|none|<i><b>Fig.1:</b>(U=Uncut C=Cut) Restriction Digestion of b-arrestin-TEV protease with SlaI, Expected bands : 2014 bp + 892 bp + 760 bp + 394 bp, Positive Result: green arrows</i>]]
  
  
 
===Sequence and Features===
 
===Sequence and Features===
 
<partinfo>BBa_K3505024 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3505024 SequenceAndFeatures</partinfo>

Revision as of 15:09, 26 October 2020


B-arrestin:TEV Protease GB compatible with B2-B5

long


Usage and Biology

Design Notes

The coding sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is presents in pUPD2 and has overhangs compatible for Golden Braid cloning. The CDS has position B2-B5.

File:T--Thessaly--GB-CCAT-GCTT.jpeg .jpeg
Figure 1.The overhangs of this part in the Golden Braid Grammar.


Figure 2. The level 0 module : pupd2-b-arrestin-TEV protease (illustration from SnapGene)

Verification of Cloning

Fig.1:(U=Uncut C=Cut) Restriction Digestion of b-arrestin-TEV protease with SlaI, Expected bands : 2014 bp + 892 bp + 760 bp + 394 bp, Positive Result: green arrows


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 1611
    Illegal PstI site found at 303
    Illegal PstI site found at 390
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 1611
    Illegal PstI site found at 303
    Illegal PstI site found at 390
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 203
    Illegal XhoI site found at 600
    Illegal XhoI site found at 994
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 1611
    Illegal PstI site found at 303
    Illegal PstI site found at 390
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 1611
    Illegal PstI site found at 303
    Illegal PstI site found at 390
    Illegal AgeI site found at 586
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1545
    Illegal SapI.rc site found at 1893